CccS and CccP are Involved in Construction of Cell Surface Components in the Cyanobacterium Synechocystis sp. strain PCC 6803

Yoshimura, Hidehisa; Kaneko, Yasuko; Ehira, Shigeki; Yoshihara, Shizue; Ikeuchi, Masahiko; Ohmori, Masayuki

doi:10.1093/pcp/pcq081

Cited by 19 publications

(23 citation statements)

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“…To tackle these questions we use here surface plasmon resonance (SPR) to detect in a highly sensitive manner the binding of proteins to sensorchip-bound DNA. We have utilized regulator box-containing promoter fragments from three Synechocystis genes, two of them, glnA and glnN, encoding glutamine synthetases types I and III [20,21], exhibiting a canonical and a non-canonical NtcA-binding site [22], respectively, and the third one, cccS, being a well characterized SYCRP1-regulation target [9,23] (Fig. 1, bottom table).…”

Section: Introductionmentioning

confidence: 99%

SPR analysis of promoter binding of Synechocystis PCC6803 transcription factors NtcA and CRP suggests cross‐talk and sheds light on regulation by effector molecules

2014

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Edited by Miguel De la RosaKeywords: Protein-protein interaction PipX protein 2-Oxoglutarate Cyanobacteria Catabolite gene activator protein Nitrogen metabolism a b s t r a c t Surface plasmon resonance monitoring of the binding of transcription factors cAMP receptor protein (CRP) and nitrogen control factor of cyanobacteria (NtcA) from Synechocystis sp. PCC6803 to promoter fragments of glnA, glnN (NtcA regulon) and cccS (CRP regulon), revealed exclusive CRP binding to cccS, whereas NtcA was bound to all three promoters with different affinities, which were strongly increased by the NtcA activator 2-oxoglutarate. Effective NtcA affinity for 2-oxoglutarate varied with the promoter. High-affinity promoters and the NtcA-coactivating protein PII-interacting protein X (PipX) increased NtcA affinity towards 2-oxoglutarate, suggesting PipX-stabilization of the 2-oxoglutarate-bound NtcA conformation. PipX binding to NtcA required 2-oxoglutarate and was much tighter (K d % 85 nM) than to the PipX-sequestering PII protein. NtcA appears to require more strongly PipX and 2-oxoglutarate (2OG) for estimulating gene expression at promoters having ''imperfect'' NtcA binding sites.

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Section: Introductionmentioning

confidence: 99%

SPR analysis of promoter binding of Synechocystis PCC6803 transcription factors NtcA and CRP suggests cross‐talk and sheds light on regulation by effector molecules

2014

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“…The CccP (Slr1668) and CccS (Slr1667) proteins involved in construction of cell surface components and in motility processes (Yoshimura et al . ), increased in high Cu 2+ but returned to normal levels upon the metal ion depletion. Enzymes engaged in peptidoglycan biosynthesis, MurA (Slr0017), MurG (Slr1656), GlmU (Sll0899) and LpdX (Slr0776), demonstrated various dynamics.…”

Section: Resultsmentioning

confidence: 86%

Global proteome response ofSynechocystis6803 to extreme copper environments applied to control the activity of the induciblepetJpromoter

et al. 2019

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Aims Cyanobacteria are prokaryotes performing oxygenic photosynthesis, and they can be engineered to harness solar energy for production of commodity and high‐value chemicals by means of synthetic biology. The Cu2+‐regulated petJ promoter (PpetJ), which controls the expression of the endogenous cytochrome c553, can be used for expression of foreign products in Synechocystis 6803. We aimed to disclose potential bottlenecks in application of the PpetJ in synthetic biology approaches. Methods and Results Quantitative label‐free mass spectrometry revealed global proteome changes which occurred during nutrient conditions which repress or activate of PpetJ in Synechocystis 6803. Conclusions Some irreversible proteome alterations were discovered due to the copper stress, including downregulation of the ribosomal proteins, significant changes in protein amounts of the cell surface layer and the outer and inner membranes. Significance and Impact of the Study This study revealed limitations in the use of PpetJ for biotechnological applications.

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“…The cccS gene product localizes at the cell membrane and contributes to cell surface construction of membrane proteins and pilins (52).…”

Section: Resultsmentioning

confidence: 99%

Using Transcriptomics To Improve Butanol Tolerance of Synechocystis sp. Strain PCC 6803

Anfelt

Hallström

Nielsen

et al. 2013

Appl Environ Microbiol

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c Cyanobacteria are emerging as promising hosts for production of advanced biofuels such as n-butanol and alkanes. However, cyanobacteria suffer from the same product inhibition problems as those that plague other microbial biofuel hosts. High concentrations of butanol severely reduce growth, and even small amounts can negatively affect metabolic processes. An understanding of how cyanobacteria are affected by their biofuel product can enable identification of engineering strategies for improving their tolerance. Here we used transcriptome sequencing (RNA-Seq) to assess the transcriptome response of Synechocystis sp. strain PCC 6803 to two concentrations of exogenous n-butanol. Approximately 80 transcripts were differentially expressed at 40 mg/liter butanol, and 280 transcripts were different at 1 g/liter butanol. Our results suggest a compromised cell membrane, impaired photosynthetic electron transport, and reduced biosynthesis. Accumulation of intracellular reactive oxygen species (ROS) scaled with butanol concentration. Using the physiology and transcriptomics data, we selected several genes for overexpression in an attempt to improve butanol tolerance. We found that overexpression of several proteins, notably, the small heat shock protein HspA, improved tolerance to butanol. Transcriptomics-guided engineering created more solventtolerant cyanobacteria strains that could be the foundation for a more productive biofuel host. C yanobacteria are attractive biochemical production platforms due to their minimal nutrient requirements, ease of genetic manipulation, and wide natural diversity. Several model cyanobacteria have been engineered as hosts for production of chemicals and biofuels such as hydrogen (1), ethanol (2), isobutanol (3, 4), and n-butanol (5). The modified cyanobacterial strains contained enzymes from fermentative organisms, such as Clostridium, Lactococcus, and Zymomonas species. The alcohols were produced at levels (up to 450 mg/liter) that were detectable but well below the titers of 10 to 20 g/liter achieved by industrial Clostridium and Saccharomyces strains (6). In addition to restrictions on metabolic flux (7,8), hosts may suffer from product inhibition even at low levels of product (9). Biofuel hosts that are more tolerant to the product may be more efficient producers even at levels below toxicity (10, 11), though this phenomenon is not universal (12).The deleterious effects of solvents on a wide range of microbes have been reported and recently reviewed (13). Solvents compromise the cell membrane and alter membrane fluidity (14). A compromised cell membrane can leak metabolites, resulting in loss of the transmembrane electrochemical gradient and hence of the proton-motive force. Reactive oxygen species (ROS) may accumulate as the cell modulates respiration to recover lost ATP (9, 15). These effects have been observed for bacteria, archaea, and yeast. Recent studies have begun to describe the complex response of cyanobacteria to solvents such as ethanol (16), n-butanol (17), and hexane (...

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CccS and CccP are Involved in Construction of Cell Surface Components in the Cyanobacterium Synechocystis sp. strain PCC 6803

Cited by 19 publications

References 50 publications

SPR analysis of promoter binding of Synechocystis PCC6803 transcription factors NtcA and CRP suggests cross‐talk and sheds light on regulation by effector molecules

SPR analysis of promoter binding of Synechocystis PCC6803 transcription factors NtcA and CRP suggests cross‐talk and sheds light on regulation by effector molecules

Global proteome response ofSynechocystis6803 to extreme copper environments applied to control the activity of the induciblepetJpromoter

Using Transcriptomics To Improve Butanol Tolerance of Synechocystis sp. Strain PCC 6803

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